Journal: bioRxiv

Article Title: SEASONAL PM 2.5 DIFFERENTIALLY REGULATES JAK2/STAT3 SIGNALING IN RURAL AND URBAN COHORTS

doi: 10.1101/2025.09.25.678452

Figure Lengend Snippet: a. Transcript profile of PIAS2 and SOCS2 in airway cells of RU and UR groups during monsoon and winter. b. Transcript profile of PIAS2 and SOCS2 in leucocytes of RU and UR groups during monsoon and winter. c. Representative Western blots of PIAS2 and SOCS2 during both the seasons of RU and UR groups. d. Mean band intensities in airway cells of RU and UR groups. e. Representative Western blots of PIAS2 and SOCS2 during both the seasons of RU and UR groups. f. Mean band intensities in leucocytes of RU and UR groups. [ Abbreviations: LMER, Linear Mixed Effect Regression; MLR, Multiple Linear Regression; PIAS2, Protein Inhibitor of Activated STAT2; SOCS2, Suppressor of Cytokine Signaling 2; RU, Rural; UR, Urban.]

Article Snippet: Following transfer, membranes were blocked with 5% BSA and incubated overnight at 4 °C with primary antibodies against PIAS2 (Cat#AF0273) and SOCS2 (Cat#DF8133) (Affinity Biosciences, Cincinnati, OH, USA) and β-actin (Cat#AC026) (Abclonal, Woburn, MA, USA).

Techniques: Western Blot

Journal: bioRxiv

Article Title: Functional redundancy in chicken ANP32A mediates species-specific support of avian influenza virus polymerase

doi: 10.1101/2025.03.25.645163

Figure Lengend Snippet: (A) Ni 2+ -NTA beads affinity pull-down assay demonstrating the SUMO modification of chANP32A with SUMO1, SUMO2, or SUMO3 in HEK293T cells. ( B ) Ni 2+ -NTA beads affinity pull-down assay results showing that TAK-981 treatment abolished chANP32A SUMOylation in HEK293T cells. HEK293T cells were transfected with the indicated plasmids. Twenty-four hours post-transfection, TAK-981 was added and incubated for an additional 24 hours prior to cell collection for the SUMOylation assay ( C, D ) Ni 2+ -NTA beads affinity pull-down assay revealing that overexpression of SENP1 or SENP2 inhibited chANP32A SUMOylation mediated by SUMO1 (C) or SUMO2 (D) in HEK293T cells. ( E ) Immunofluorescence staining analysis of co-localization between chANP32A-V5 and Flag-SENP1 or Flag-SENP2. Scale bar: 10 μm. ( F ) Co-IP experiments demonstrating the interaction between SENP1 and chANP32A in HEK293T cells. ( G ) Proximity ligation assay (PLA) confirming the interaction between endogenous SENP1 and chANP32A-Flag in HEK293T cells stably expressing chANP32A-Flag. Scale bar: 10 μm. ( H ) Ni 2+ -NTA beads affinity pull-down assay results showing enhanced chANP32A SUMOylation upon knockdown of endogenous SENP1 in HEK293T cells. ( I ) Ni 2+ -NTA beads affinity pull-down assay indicating that chANP32A SUMOylation was enhanced by overexpression of PIAS2α in HEK293T cells. ( J ) Immunofluorescence staining analysis of co-localization between chANP32A-V5 and Flag-PIAS2α. Scale bar: 20 μm. ( K ) Co-IP experiments confirming the interaction between PIAS2α and chANP32A in HEK293T cells. ( L ) PLA confirming the interaction between endogenous PIAS2 and chANP32A-Flag in HEK293T cells stably expressing chANP32A-Flag. Scale bar: 10 μm. ( M ) Ni 2+ -NTA beads affinity pull-down assay results showing chANP32A SUMOylation in HEK293T cells was notably reduced upon knockdown of PIAS2 in HEK293T cells. All the western blot experiments were independently repeated three times with consistent results.

Article Snippet: The antibodies utilized in this study included: rabbit anti-Flag (Sigma-Aldrich, F7425), rabbit anti-HA (Sigma-Aldrich, H6908), rabbit anti-ACTB (Abclonal, AC026), mouse anti-ACTB (Abclonal, AC004), rabbit anti-Myc (Abclonal, AE070), mouse anti-His (Proteintech, 66005-1-Ig), rabbit anti-V5 (Proteintech, 14440-1-AP), rabbit anti-SENP1 (Proteintech, 25349-1-AP), rabbit anti-PIAS2 (Proteintech, 16074-1-AP), rabbit anti-influenza A virus PB2 (GeneTex, GTX125926), rabbit anti-influenza A virus PA (GeneTex, GTX118991), mouse anti-influenza A virus PA (produced in our laboratory, 1:5000 for WB), DyLight 800 labeled Anti-Mouse IgG (H+L) Antibody (KPL, 5230-0415) and DyLight 680-labeled Anti-Rabbit IgG (H+L) Antibody (KPL, 5230-0402).

Techniques: Pull Down Assay, Modification, Transfection, Incubation, Over Expression, Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Proximity Ligation Assay, Stable Transfection, Expressing, Knockdown, Western Blot

Journal: Scientific reports

Article Title: Chromosome 8q24 amplification associated with human hepatocellular carcinoma predicts MYC/ZEB1/MIZ1 transcriptional regulation.

doi: 10.1038/s41598-024-75219-1

Figure Lengend Snippet: Fig. 5. Patient survival is decreased in those with c-myc amplification, high MYC, ZEB1 and MIZ1 nuclear only expression. Patient survival from the time of diagnosis with respect to tumor grade at the time of diagnosis was retrieved from patient records. The percent survival was calculated and plotted for individuals diagnosed with grade III (A) or grade IV (B) HCC.

Article Snippet: Cells were labeled with rabbit polyclonal antibodies against MYC (Cell Signaling Technology, Danvers, MA), MIZ1, ZEB1, E-cadherin or vimentin (all four from Proteintech, Rosemont, IL).

Techniques: Amplification, Expressing, Biomarker Discovery

Journal: Scientific reports

Article Title: Chromosome 8q24 amplification associated with human hepatocellular carcinoma predicts MYC/ZEB1/MIZ1 transcriptional regulation.

doi: 10.1038/s41598-024-75219-1

Figure Lengend Snippet: Fig. 8. EMT and MET in HCC cells are dynamic processes. A, More epithelial-like carcinoma cells have highest drug sensitivity, are nonproliferative and respond to apoptotic signaling whereas more mesenchymal- like cells are more proliferative, have highest drug resistance, invasion and immune evasion. A hybrid EMT state provides a plasticity window where cells can display stemness, can initiate tumor formation and can adapt to changing environments. B, Our proposed scenario for transcriptional dysregulation in HCC. In the normal polarized hepatocyte, high levels of MIZ1 are expressed (promoting polarity) whereas low levels of MYC and ZEB1 are expressed (reflecting decreased proliferation). Upon hepatic injury and the development of cirrhosis, hepatocytes become hyperplastic, then dysplastic, and as the invasive phenotype is acquired, increased chromosome instability at grades III and IV HCC is observed exemplified by c-myc amplification and increased MYC. The high Myc leads to MIZ1 transcriptional repression and loss of the epithelial phenotype. Concomitantly, the high MYC activates ZEB1 expression driving cells through EMT by increasing mesenchymal/stemness while repressing the epithelial phenotype. We further propose that as cells metastasize and colonize at secondary sites, epithelial traits are partially reacquired through the inactivation of ZEB1/MYC and reactivation of MIZ1.

Article Snippet: Cells were labeled with rabbit polyclonal antibodies against MYC (Cell Signaling Technology, Danvers, MA), MIZ1, ZEB1, E-cadherin or vimentin (all four from Proteintech, Rosemont, IL).

Techniques: Amplification, Expressing

Journal: Journal of Dental Sciences

Article Title: Protein inhibitor of activated signal transducer and activator of transcription 2 is an oncoprotein in oral squamous cell carcinoma and related to cigarette smoking - An in vitro study

doi: 10.1016/j.jds.2024.07.013

Figure Lengend Snippet: Overexpression of PIAS2 in oral cancer increases epithelial-mesenchymal transition. (A) Endogenous protein levels in oral cancer cell lines (YD38, OECM1, SAS, and SCC25). The expression of PIAS2 protein in YD38 and OECM1 (gingiva squamous cell carcinoma), and SAS and SCC25 (tongue oral squamous carcinoma cell lines) was examined using the Western blot method. (B) Quantitative results of the PIAS2 Western blot analysis. (C) PIAS2 expression in YD38 and SAS cell lines was increased by transfecting with 1 μg of pCMV Tag2A-PIAS2 in a six-well plate. After 48 h of incubation, mRNA and protein were extracted for analysis. Changes in PIAS2 mRNA levels were analyzed using qRT-PCR. (D) Changes in PIAS2 protein levels were assessed by Western blot. (E) PIAS2 expression affects EMT protein levels in oral cancer cell lines. In a six-well plate, 1 μg of PIAS2 plasmid was used to overexpress PIAS2 in YD38 and SAS cell lines. After 48 h of incubation, protein was extracted. Changes in mesenchymal marker proteins (Fibronectin) and epithelial marker proteins (E-cadherin) were assessed by Western blot. Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. PIAS2, Protein inhibitor of activated signal transducer and activator of transcription 2; EMT, epithelial-mesenchymal transition; SEM, standard error of the mean.

Article Snippet: The primary antibodies used were as follows: PIAS2 (Santa Cruz, sc-166494, Dallas, TX, USA), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell signaling, 5174, Danvers, MA, USA), E-cadherin (Cell Signaling, 14472), Fibronectin (Abcam, ab32419, Cambridge, MA, USA), Anti-rabbit or anti-mouse secondary antibodies were purchased from Jackson Immuno Research Laboratories (West Grove, PA, USA). α-Tubulin and Lamin B1 antibodies were obtained from Proteintech (Wuhan, China).

Techniques: Over Expression, Expressing, Western Blot, Incubation, Quantitative RT-PCR, Plasmid Preparation, Marker

Journal: Journal of Dental Sciences

Article Title: Protein inhibitor of activated signal transducer and activator of transcription 2 is an oncoprotein in oral squamous cell carcinoma and related to cigarette smoking - An in vitro study

doi: 10.1016/j.jds.2024.07.013

Figure Lengend Snippet: Overexpression of PIAS2 increases migration, invasion, and proliferation of human oral cancer cells. (A) Migratory ability was evaluated using a wound-healing assay. Representative images were taken at 0 and 12 h after wound scratching. Wound closure was quantified using ImageJ software. (B) Quantitative results of the wound-healing assay. (C) Invasion ability was evaluated using a transwell invasion assay. OSCC cells were treated as indicated and allowed to invade through matrigel-coated inserts for 24 h. PI-stained cells were quantified using ImageJ software. (D) Quantitative results of the invasion assay. (E) Proliferation ability was evaluated using a colony formation assay. 1000 cells were plated in six-well plates and incubated for 7 days. Crystal violet-stained colonies were quantified using ImageJ software. (F) Quantitative results of the colony formation assay. Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. PIAS2, Protein inhibitor of activated signal transducer and activator of transcription 2; OSCC, Oral squamous cell carcinoma; SEM, standard error of the mean.

Article Snippet: The primary antibodies used were as follows: PIAS2 (Santa Cruz, sc-166494, Dallas, TX, USA), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell signaling, 5174, Danvers, MA, USA), E-cadherin (Cell Signaling, 14472), Fibronectin (Abcam, ab32419, Cambridge, MA, USA), Anti-rabbit or anti-mouse secondary antibodies were purchased from Jackson Immuno Research Laboratories (West Grove, PA, USA). α-Tubulin and Lamin B1 antibodies were obtained from Proteintech (Wuhan, China).

Techniques: Over Expression, Migration, Wound Healing Assay, Software, Transwell Invasion Assay, Staining, Invasion Assay, Colony Assay, Incubation

Journal: Journal of Dental Sciences

Article Title: Protein inhibitor of activated signal transducer and activator of transcription 2 is an oncoprotein in oral squamous cell carcinoma and related to cigarette smoking - An in vitro study

doi: 10.1016/j.jds.2024.07.013

Figure Lengend Snippet: Knockdown of PIAS2 in oral cancer reverses epithelial-mesenchymal transition. (A) PIAS2 expression in OECM1 and SCC25 cell lines was decreased by transfecting with 10 nM siPIAS2 in a six-well plate. After 48 h of incubation, mRNA and protein were extracted for analysis. Changes in PIAS2 mRNA levels were analyzed using qRT-PCR. (B) Changes in PIAS2 protein levels were assessed by Western blot. (C) PIAS2 expression affects EMT protein levels in oral cancer cell lines. In a six-well plate, 10 nM siPIAS2 was used to reduce PIAS2 expression in OECM1 and SCC25 cell lines. After 48 h of incubation, protein was extracted. Changes in mesenchymal marker proteins (Fibronectin) and epithelial marker proteins (E-cadherin) were assessed by Western blot. Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. PIAS2, Protein inhibitor of activated signal transducer and activator of transcription 2; EMT, epithelial-mesenchymal transition; SEM, standard error of the mean.

Article Snippet: The primary antibodies used were as follows: PIAS2 (Santa Cruz, sc-166494, Dallas, TX, USA), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell signaling, 5174, Danvers, MA, USA), E-cadherin (Cell Signaling, 14472), Fibronectin (Abcam, ab32419, Cambridge, MA, USA), Anti-rabbit or anti-mouse secondary antibodies were purchased from Jackson Immuno Research Laboratories (West Grove, PA, USA). α-Tubulin and Lamin B1 antibodies were obtained from Proteintech (Wuhan, China).

Techniques: Knockdown, Expressing, Incubation, Quantitative RT-PCR, Western Blot, Marker

Journal: Journal of Dental Sciences

Article Title: Protein inhibitor of activated signal transducer and activator of transcription 2 is an oncoprotein in oral squamous cell carcinoma and related to cigarette smoking - An in vitro study

doi: 10.1016/j.jds.2024.07.013

Figure Lengend Snippet: Knockdown of PIAS2 decreases malignant behaviors of human oral cancer cells. (A) Migratory ability was evaluated using a wound-healing assay. Representative images were taken at 0 and 12 h after wound scratching. Wound closure was quantified using ImageJ software. (B) Quantitative results of the wound-healing assay. (C) Invasion ability was evaluated using a transwell invasion assay. OSCC cells were treated as indicated and allowed to invade through matrigel-coated inserts for 24 h. PI-stained cells were quantified using ImageJ software. (D) Quantitative results of the invasion assay. (E) Proliferation ability was evaluated using a colony formation assay. 1000 cells were plated in six-well plates and incubated for 7 days. Crystal violet-stained colonies were quantified using ImageJ software. (F) Quantitative results of the colony formation assay. Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. PIAS2, Protein inhibitor of activated signal transducer and activator of transcription 2; OSCC, Oral squamous cell carcinoma; SEM, standard error of the mean.

Article Snippet: The primary antibodies used were as follows: PIAS2 (Santa Cruz, sc-166494, Dallas, TX, USA), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell signaling, 5174, Danvers, MA, USA), E-cadherin (Cell Signaling, 14472), Fibronectin (Abcam, ab32419, Cambridge, MA, USA), Anti-rabbit or anti-mouse secondary antibodies were purchased from Jackson Immuno Research Laboratories (West Grove, PA, USA). α-Tubulin and Lamin B1 antibodies were obtained from Proteintech (Wuhan, China).

Techniques: Knockdown, Wound Healing Assay, Software, Transwell Invasion Assay, Staining, Invasion Assay, Colony Assay, Incubation

Journal: Journal of Dental Sciences

Article Title: Protein inhibitor of activated signal transducer and activator of transcription 2 is an oncoprotein in oral squamous cell carcinoma and related to cigarette smoking - An in vitro study

doi: 10.1016/j.jds.2024.07.013

Figure Lengend Snippet: CSC treatment increases PIAS2 expression in oral cancer cells. Dose-dependent changes in PIAS2 expression were observed after treatment with CSC in (A) YD38 and (B) SAS cells. CSC, cigarette smoke condensate; PIAS2, Protein inhibitor of activated signal transducer and activator of transcription 2.

Article Snippet: The primary antibodies used were as follows: PIAS2 (Santa Cruz, sc-166494, Dallas, TX, USA), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell signaling, 5174, Danvers, MA, USA), E-cadherin (Cell Signaling, 14472), Fibronectin (Abcam, ab32419, Cambridge, MA, USA), Anti-rabbit or anti-mouse secondary antibodies were purchased from Jackson Immuno Research Laboratories (West Grove, PA, USA). α-Tubulin and Lamin B1 antibodies were obtained from Proteintech (Wuhan, China).

Techniques: Expressing